A new generation of sequencing technologies is emerging that will decrease the cost of many standard sequencing applications and change the manner in which researchers approach fundamental questions of biomedical importance, many of which were formerly out of the reach of most investigators. The objective of this proposal is to create a new high-throughput screening (HTS) facility at Stanford University based on the first commercialized ultra-low cost sequencing technology: the "Genome Sequencer 20 System" developed by 454 Life Sciences. The 454 Sequencing System performs highly parallel emulsion-based clonal amplification and pyrosequencing of 200,000 DNA templates, routinely producing a minimum of 20 million bp in a four-hour period. A HTS facility at the Stanford Genome Center will not only facilitate ongoing research projects, but will also attract many additional researchers to the Center, creating synergistic interactions among experts in sequencing, functional genomics, translational research, and bio-informatics. Eight NIH-funded researchers, including four major users, have proposed projects that would not be possible without the high throughput of the 454 sequencing system. The specific projects outlined in this proposal include microbial genome sequencing, unbiased explorations of human microbial diversity (bacterial and viral), and HIV-1 quasispecies analyses. These projects, however, represent only a few of the many potential applications of the 454 Sequencing System. The scientific oversight committee will foster and cultivate additional applications, including analyses of gene expression and alternative splicing, human genotype-phenotype correlations, and mutational pathways associated with the development and spread of malignancy. Finally, this proposal is unique in the fact that the principal investigator invented pyrosequencing and has extensive experience in its use. As a result, Stanford investigators will have immediate access to expert assistance in experiment design and data analysis. The diverse array of research projects supported by the HTS facility will spark the development of new applications, accelerating the Center's technology development program and opening a new horizon in biomedical research at Stanford. [unreadable] [unreadable] [unreadable]